Pharmaceutical preparation for subcutaneous administration, comprising infliximab and hyaluronidase

Abstract

The present invention relates to a pharmaceutical preparation for subcutaneous administration, comprising infliximab and native human recombinant hyaluronidase PH20.

Description

Subcutaneous pharmaceutical preparation containing infliximab and hyaluronidase

[0001] The present invention relates to a pharmaceutical preparation for subcutaneous administration comprising infliximab and hyaluronidase.

[0002] Infliximab is a chimeric monoclonal antibody that acts as a tumor necrosis factor-α (TNF-α) inhibitor. Conventional formulations containing this antibody are manufactured as lyophilized powders, which are reconstituted and diluted for intravenous injection according to the appropriate dosage and route of administration for each disease. However, these conventional methods are costly, cumbersome, and require frequent administration, leading to patient discomfort, resistance, and side effects. Furthermore, they are limited to only medically trained individuals.

[0003] To address the issues associated with intravenous administration, a subcutaneous formulation containing infliximab alone has been developed, resulting in improved dosing options being marketed. However, this commercially available subcutaneous formulation of infliximab alone has the limitation of requiring high doses of infliximab every two weeks. Therefore, research is needed to reduce the infliximab content or dose per dose, as well as to extend the dosing interval or reduce the number of doses.

[0004] The present invention provides a pharmaceutical preparation for subcutaneous administration, including infliximab and natural human recombinant hyaluronidase PH20.

[0005] However, the technical problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

[0006] The present invention provides a pharmaceutical preparation for subcutaneous administration comprising infliximab and a natural human recombinant hyaluronidase PH20.

[0007] The above natural human recombinant hyaluronidase PH20 may be used for the diffusion of the above infliximab.

[0008] The content of the above infliximab may be 30 mg / mL to 240 mg / mL, and the content of the above natural human recombinant hyaluronidase PH20 may be 1,000 unit / mL to 10,000 unit / mL.

[0009] When the above pharmaceutical preparation for subcutaneous administration is administered subcutaneously, the single administration dose of infliximab may be 30 mg to 600 mg.

[0010] When the above pharmaceutical preparation for subcutaneous administration is administered subcutaneously, the area under the concentration-time curve (AUC) of infliximab in plasma may be 120% or more compared to when the preparation containing infliximab alone is subcutaneously administered at the same dose.

[0011] As a buffer, a phosphate buffer or a succinic acid buffer may be additionally included.

[0012] As a buffer, it may additionally contain 1 mM to 20 mM succinic acid.

[0013] The above pharmaceutical preparation for subcutaneous administration may exclude salt.

[0014] As a surfactant, a polysorbate-based material; as a sugar or its derivative, at least one selected from the group consisting of sorbitol, mannitol, trehalose, and sucrose; and as a viscosity-reducing agent, methionine or arginine may be additionally included.

[0015] As a surfactant, it may further include 0.05 mg / mL to 5 mg / mL of polysorbate 80; as a sugar or its derivative, 10 mg / mL to 100 mg / mL of sorbitol; and as a viscosity-reducing agent, 0.5 mg / mL to 20 mg / mL of methionine or arginine.

[0016] The above pharmaceutical preparation for subcutaneous administration may have a viscosity of 1.0 mPa / s to 12.0 mPa / s under temperature conditions of 25°C.

[0017] The pharmaceutical preparation for subcutaneous administration according to the present invention comprises infliximab and a natural human recombinant hyaluronidase PH20, and has the advantage of effectively diffusing infliximab by combining the natural human recombinant hyaluronidase PH20 with infliximab.

[0018] Therefore, the present invention has the advantage of i) showing the same or greater efficacy as the commercially available infliximab single subcutaneous formulation while lowering the content or single administration dose of infliximab, or ii) increasing the administration volume and single administration dose while maintaining the content of infliximab, thereby increasing the blood drug exposure amount, thereby extending the administration interval or reducing the number of administrations. Meanwhile, the commercially available infliximab single subcutaneous formulation is used after the first two intravenous injections, but according to the present invention, iii) it has the effect of being able to be replaced with the pharmaceutical preparation for subcutaneous administration according to the present invention from the first two intravenous injections.

[0019] Moreover, the pharmaceutical preparation for subcutaneous administration according to the present invention can secure a viscosity suitable for subcutaneous administration while maintaining high stability of infliximab and natural human recombinant hyaluronidase PH20 enzyme activity by applying a phosphate buffer or a succinic acid buffer (particularly, succinic acid) as a buffer, and thus can be utilized as a subcutaneous administration formulation.

[0020] Figure 1 is a graph showing the results of analyzing the change in the plasma concentration of infliximab over time by subcutaneously injecting once the subcutaneous administration formulation according to Comparative Example 1 and Examples 1 to 3 into SD rats and isolating the plasma for up to 28 days.

[0021] Figure 2 is a graph showing the results of analyzing the change in the plasma concentration of infliximab over time by isolating plasma for up to 28 days after subcutaneously injecting the subcutaneous administration formulation according to Comparative Example 2 and Examples 8 and 16 once into SD rats.

[0022] Figure 3 is a graph showing the results of analyzing the change in the plasma concentration of infliximab over time by isolating plasma for up to 28 days after subcutaneously injecting once the subcutaneous administration formulation according to Comparative Example 2 and Examples 8, 17 to 19 into SD rats.

[0023] Figure 4 is a graph showing the results of analyzing the change in the plasma concentration of infliximab over time by isolating plasma for up to 28 days after intravenous administration formulation according to Comparative Example 3 and subcutaneous administration formulation according to Examples 17 and 18 were administered intravenously / subcutaneously once to SD rats.

[0024] The present inventors conducted research to reduce the single dose of infliximab or increase the dosing interval when manufacturing a pharmaceutical formulation for subcutaneous administration of infliximab. As a result, they developed a subcutaneous formulation that combines infliximab with natural human recombinant hyaluronidase PH20 and a specific buffer. Accordingly, they confirmed that this formulation not only effectively disperses infliximab but also secures both viscosity characteristics and stability desirable for subcutaneous administration, thereby completing the present invention.

[0025]

[0026] Hereinafter, the present invention will be described in detail.

[0027]

[0028] Subcutaneous pharmaceutical preparation containing infliximab and hyaluronidase

[0029]

[0030] The present invention provides a pharmaceutical preparation for subcutaneous administration comprising infliximab and a natural human recombinant hyaluronidase PH20.

[0031]

[0032] First, the pharmaceutical preparation for subcutaneous administration according to the present invention includes infliximab.

[0033] Infliximab, described above, is a chimeric monoclonal antibody that acts as a tumor necrosis factor (TNF)-α inhibitor. It blocks TNF-α from binding to its receptor and activates it, causing the breakdown of TNF-α-producing cells. Consequently, it reduces inflammation caused by excessive TNF-α activity and exerts immunosuppressive effects. Infliximab is an antibody therapeutic (monoclonal antibody) created using genetic recombination technology to specifically bind to TNF-α. Unlike other chemically synthesized drugs, it is a biopharmaceutical derived from living organisms.

[0034] The content of the above infliximab may be 30 mg / mL to 240 mg / mL, preferably 50 mg / mL to 150 mg / mL, but is not limited thereto. In this case, if the content of the above infliximab is too low, there is a problem that the effective efficacy is not sufficiently expressed, and if the content of the above infliximab is too high, it may be difficult to implement it as a subcutaneous administration formulation.

[0035] When the above pharmaceutical formulation for subcutaneous administration is administered subcutaneously, the single dose of infliximab may be 30 mg to 600 mg. To increase the single dose of infliximab, the content of infliximab itself may be increased, but the administration volume may also be increased while maintaining the content for implementation in a subcutaneous administration formulation. In this case, the administration volume may be 0.5 mL to 10 mL, and preferably 1 mL to 5 mL.

[0036] According to the present invention, due to the diffusion action of the natural human recombinant hyaluronidase PH20 described below, when the above-described subcutaneous pharmaceutical preparation is administered subcutaneously, the area under the concentration-time curve (AUC) of infliximab in the plasma may be 120% or more, compared to when the preparation containing infliximab alone is subcutaneously administered at the same dose. Meanwhile, the maximum plasma concentration (C) of infliximab max ) value can be greater than 110%.

[0037] In particular, due to the diffusion action of the natural human recombinant hyaluronidase PH20 described below, i) it is possible to show efficacy equivalent to or greater than that of a commercially available infliximab single subcutaneous formulation while reducing the content or single dose of infliximab. For example, when the single dose of infliximab is reduced by 25% in subcutaneously administering the above pharmaceutical formulation for subcutaneous administration, the area under the concentration-time curve of infliximab in the plasma (AUC) value may be about 100% or more compared to when a formulation containing infliximab alone is subcutaneously administered. Meanwhile, the maximum plasma concentration (C) of infliximab max ) value may be about 90% or more. Accordingly, in the present invention, in case of i), the content of infliximab can be kept relatively low, such as 30 mg / mL to less than 100 mg / mL (preferably, 50 mg / mL to less than 100 mg / mL). Alternatively, the single administration dose of infliximab can be kept relatively low, such as 30 mg to less than 75 mg.

[0038] Alternatively, ⅱ) By increasing the administration volume and single administration dose while maintaining the content of infliximab, there is an advantage in that the administration interval can be extended or the number of administrations can be reduced by increasing the blood drug exposure amount. As another example, when the single administration dose of infliximab is increased up to 4 times in subcutaneous administration of the above-mentioned pharmaceutical preparation for subcutaneous administration, the area under the concentration-time curve of infliximab in the plasma (The area under the concentration, AUC) value can be about 500% or more compared to when a preparation containing infliximab alone is subcutaneously administered. Meanwhile, the maximum plasma concentration (C) of infliximab max ) value may also be about 500% or more. Accordingly, in the present invention, in the case of ⅱ), while maintaining the content of infliximab at 100 mg / mL to 240 mg / mL (preferably, 100 mg / mL to 150 mg / mL), the single administration dose of infliximab can be maintained relatively high, such as 75 mg to 600 mg.

[0039] Meanwhile, in the case of the commercially available subcutaneous administration formulation of infliximab alone, it is used after the first two intravenous injections. However, according to the present invention, iii) it has the effect of being able to be replaced with the subcutaneous administration pharmaceutical preparation according to the present invention from the first two intravenous injections. In another example, when the pharmaceutical preparation for subcutaneous administration is subcutaneously administered, if the single dose of infliximab is 168 mg or more, the area under the concentration-time curve of infliximab in the plasma (AUC) may be about 90% or more compared to the case of intravenous administration of a preparation containing infliximab alone. Meanwhile, the maximum plasma concentration (C) of infliximab max) value may be 50% or more. Accordingly, in the present invention, in the first two cases in ⅲ), it is necessary to maintain the single administration dose of infliximab at a high dose, such as 90 mg to 600 mg, while maintaining the content of infliximab at 100 mg / mL to 240 mg / mL (preferably, 100 mg / mL to 150 mg / mL). From the third time thereafter, the single administration dose of infliximab may be maintained at a relatively low dose, such as 30 mg to less than 90 mg (preferably, 60 mg to less than 90 mg).

[0040]

[0041] Next, the pharmaceutical preparation for subcutaneous administration according to the present invention comprises a natural human recombinant hyaluronidase PH20, which may be used for diffusion of infliximab.

[0042] The above natural human recombinant hyaluronidase PH20 may be a human-derived hyaluronidase PH20 known in the art, and is preferably represented by the amino acid sequence of sequence number 1, but is not limited thereto.

[0043] Unlike its PH20 variants, the above-mentioned natural human recombinant hyaluronidase PH20 possesses the same amino acid sequence and three-dimensional structure as the endogenous human PH20 protein (enzyme). Accordingly, it is less likely to act as an antigen when injected externally, thus providing the advantage of safety when administered to the human body.

[0044] The content of the natural human recombinant hyaluronidase PH20 may be 1,000 unit / mL to 10,000 unit / mL, preferably 1,000 unit / mL to 5,000 unit / mL, and more preferably 1,000 unit / mL to 4,000 unit / mL, but is not limited thereto. In this case, if the content of the natural human recombinant hyaluronidase PH20 is too low, there is a problem that the diffusion effect of infliximab is not sufficient, and if the content of the natural human recombinant hyaluronidase PH20 is too high, it may be difficult to implement it as a subcutaneous administration formulation.

[0045] Meanwhile, when the above-mentioned pharmaceutical preparation for subcutaneous administration is administered subcutaneously, the single administration dose of the natural human recombinant hyaluronidase PH20 may be 1,000 units to 10,000 units. In order to increase the single administration dose of the natural human recombinant hyaluronidase PH20, the content of the natural human recombinant hyaluronidase PH20 itself may be increased, but in order to implement it as a subcutaneous administration formulation, the administration volume may be increased while maintaining the content.

[0046]

[0047] Next, the subcutaneous pharmaceutical formulation according to the present invention may additionally include a buffer, such as a phosphate buffer or a succinic acid buffer. Such a buffer maintains the subcutaneous pharmaceutical formulation at a pH of 4.5 to 5.5, thereby enabling the stable storage of highly concentrated infliximab.

[0048] Specifically, the phosphate buffer may be phosphate-citrate, and the succinic acid buffer may be succinic acid, and the concentration thereof may be 1 mM to 20 mM, preferably 5 mM to 15 mM, but is not limited thereto.

[0049] Meanwhile, when sodium citrate is used as a buffer, although stability is maintained at a pH level, the viscosity increases excessively, resulting in a gelatinous appearance. This means that when used as a subcutaneous pharmaceutical preparation, there are limitations such as the possibility of tissue damage at the injection site, pain, and inadequate drug distribution.

[0050]

[0051] If necessary, the subcutaneous pharmaceutical formulation according to the present invention may include salts such as NaCl to increase ionic strength and stability. However, such salts may cause a significant increase in viscosity when combined with succinic acid as a specific buffer, and thus are preferably excluded, but are not limited thereto.

[0052]

[0053] Next, the pharmaceutical preparation for subcutaneous administration according to the present invention may further include a polysorbate-based substance as a surfactant; at least one selected from the group consisting of sorbitol, mannitol, trehalose, and sucrose as a sugar or derivative thereof; and methionine or arginine as a viscosity-reducing agent.

[0054] Specifically, the polysorbate-based material is intended to prevent aggregation of highly concentrated infliximab, and may be at least one selected from the group consisting of polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80, preferably 0.05 mg / mL to 5 mg / mL of polysorbate 80, more preferably 0.1 mg / mL to 1 mg / mL of polysorbate 80, but is not limited thereto. In addition, the sugar or its derivative acts as a protein stabilizer, and is intended to suppress osmotic stress and reduce viscosity, and may be at least one selected from the group consisting of sorbitol, mannitol, trehalose, and sucrose, and is preferably 10 mg / mL to 100 mg / mL of sorbitol, more preferably 30 mg / mL to 50 mg / mL of sorbitol, but is not limited thereto. In addition, the viscosity-lowering agent is also for lowering viscosity, and may be methionine or arginine, and its concentration may be 0.5 mg / mL to 20 mg / mL, and is preferably 0.5 mg / mL to 5 mg / mL, but is not limited thereto.

[0055]

[0056] The pharmaceutical preparation for subcutaneous administration according to the present invention is implemented as a formulation that additionally includes a phosphate buffer or a succinic acid buffer as a buffer, and thus can maintain a viscosity of 1.0 mPa / s to 12.0 mPa / s under a temperature condition of 25°C. In particular, by additionally including succinic acid as a specific buffer and omitting the mixing of salts, the viscosity can be maintained of 1.0 mPa / s to 6.0 mPa / s under a temperature condition of 25°C, and thus can be considered the most preferable pharmaceutical preparation for subcutaneous administration.

[0057] The term "viscosity at 25°C" in this specification refers to the dynamic viscosity, which represents the internal frictional resistance when a pharmaceutical preparation for subcutaneous administration flows. This may be measured immediately after manufacture, or may be measured after storage under refrigerated conditions (2-8°C) for less than 8 weeks (preferably less than 4 weeks). At this time, a commercial viscometer such as m-VROC II can be used to measure the viscosity.

[0058]

[0059] In addition, the pharmaceutical preparation for subcutaneous administration according to the present invention has the advantage of maintaining high stability of infliximab and natural human recombinant hyaluronidase PH20 enzyme activity under refrigerated conditions (2-8°C).

[0060]

[0061] The pharmaceutical preparation for subcutaneous administration according to the present invention can be used to treat diseases caused by TNF-α overexpression. Specifically, the diseases caused by TNF-α overexpression may include at least one selected from the group consisting of sepsis, autoimmune diseases, infectious diseases, transplantation, malignant cancer, lung disorders, intestinal disorders, and cardiac disorders, and preferably includes at least one selected from the group consisting of rheumatoid arthritis, ankylosing spondylitis, ulcerative colitis, adult Crohn's disease, pediatric Crohn's disease, psoriasis, and psoriatic arthritis, but is not limited thereto.

[0062] In addition, the pharmaceutical preparation for subcutaneous administration according to the present invention is limited to a subcutaneous injection method in which the administration method is administered into the adipose tissue of the subcutaneous tissue located under the epidermis and dermis, and can be injected once a week to once every four weeks, and can be administered once or a total of five times. Such administration intervals and frequency can be controlled by a single administration dose of infliximab and the diffusion action of the native human recombinant hyaluronidase PH20.

[0063]

[0064] Alternatively, the present invention provides a method of subcutaneously administering to a subject a pharmaceutical formulation comprising infliximab and a native human recombinant hyaluronidase PH20.

[0065] Alternatively, the present invention provides a method for preventing or treating a disease caused by TNF-α overexpression, comprising the step of subcutaneously administering to a subject a pharmaceutical formulation comprising infliximab and a native human recombinant hyaluronidase PH20. Here, the term "subject" refers to a subject in need of treatment for a disease, and more specifically, refers to a mammal such as a human or non-human primate, mouse, rat, dog, cat, horse, or cow.

[0066]

[0067] As reviewed above, the pharmaceutical preparation for subcutaneous administration according to the present invention comprises infliximab and a natural human recombinant hyaluronidase PH20, and has the advantage of effectively diffusing infliximab by combining the natural human recombinant hyaluronidase PH20 with infliximab.

[0068] Therefore, the present invention has the advantage of i) showing the same or greater efficacy as the commercially available infliximab single subcutaneous formulation while lowering the content or single administration dose of infliximab, or ii) increasing the administration volume and single administration dose while maintaining the content of infliximab, thereby increasing the blood drug exposure amount, thereby extending the administration interval or reducing the number of administrations. Meanwhile, the commercially available infliximab single subcutaneous formulation is used after the first two intravenous injections, but according to the present invention, iii) it has the effect of being able to be replaced with the pharmaceutical preparation for subcutaneous administration according to the present invention from the first two intravenous injections.

[0069] Moreover, the pharmaceutical preparation for subcutaneous administration according to the present invention can secure a viscosity suitable for subcutaneous administration while maintaining high stability of infliximab and natural human recombinant hyaluronidase PH20 enzyme activity by applying a phosphate buffer or a succinic acid buffer (particularly, succinic acid) as a buffer, and thus can be utilized as a subcutaneous administration formulation.

[0070]

[0071] Hereinafter, preferred examples are presented to aid in understanding the present invention. However, the following examples are provided solely to facilitate a better understanding of the present invention, and the scope of the present invention is not limited by the following examples.

[0072]

[0073] [Example]

[0074] Example 1-3 and Comparative Example 1: Infliximab diffusion effect depending on the addition of natural human recombinant hyaluronidase PH20

[0075] Natural human recombinant hyaluronidase PH20 (rHuPH20, product name: HyDIFFUZE) TM ) In order to confirm the diffusion effect of infliximab, a composition containing only 120 mg / mL infliximab was used as comparative example 1, and HyDIFFUZE was added to 60, 90, and 120 mg / mL infliximab. TM Compositions containing 2,000 units / mL were prepared in Examples 1-3, respectively (Table 1).

[0076] HyDIFFUZE Infliximab content / single dose TM Content / Single dose comparison Example 1 Infliximab 120 mg / mL - Infliximab 84 mg / kg - Example 1 Infliximab 60 mg / mL HyDIFFUZE TM 2,000 unit / mL infliximab 42 mg / kg HyDIFFUZE TM 1,400 units / kgExample 2 Infliximab 90 mg / mLHyDIFFUZE TM2,000 unit / mL infliximab 63 mg / kg HyDIFFUZE TM 1,400 units / kgExample 3 Infliximab 120 mg / mLHyDIFFUZE TM 2,000 unit / mL infliximab 84 mg / kg HyDIFFUZE TM 1,400 units / kg

[0077] Afterwards, a single subcutaneous injection was given to SD rats, and approximately 0.5 mL of blood was collected from the jugular vein using a disposable syringe (1 mL, 25 G) at 0 hours before administration, 2 and 7 hours after administration, and 1, 2, 3, 4, 7, 10, 14, 21, and 28 days. The blood was then placed in an EDTA tube and centrifuged at 12,000 rpm for 3 minutes to separate the plasma.

[0078] To quantitatively analyze infliximab in SD rat plasma, the antibody ELISA kit (Matriks biotek, Cat. No. INF-FD-REMI) was used. Antibody standard solutions were serially diluted 1 / 3 with Assay buffer within the calibration range of 0.41–300 ng / mL, and samples were prepared by diluting them in Assay buffer so that they could be measured within the calibration range. The experiment was performed according to the ELISA kit analysis procedure, and the absorbance at 450 nm was measured using a microplate reader. Based on the analysis results, pharmacokinetic analysis was performed using the WinNonlin program (Table 2 and Fig. 1).

[0079] Distinction AUC last (㎍·hr / mL)C max (㎍ / mL)T max (hr)t 1 / 2 (hr)CL t(mL / hr / kg)Vd(mL / kg)MRT(hr)Comparative Example 1Average62056.33331.0176.8040.461.3778.57136.43Standard Deviation8696.3120.1520.0811.240.2019.4621.31N5555555Example 1Average48378.53238.7472.0045.830.9158.96140.83Standard Deviation13372.8620.5716.9713.570.2419.9527.02N5555555Example 2Mean 64542.24313.9186.4038.500.9953.47141.06Standard deviation 9381.2241.0613.159.480.147.6122.55N5555555Example 3Mean 79448.14385.3467.2031.881.0849.69142.38Standard deviation 14737.8450.4820.084.330.1710.6111.06N5555555

[0080] As a result, the content / single dose of infliximab is equivalent and HyDIFFUZE TM In Example 3, HyDIFFUZE was added TM Compared to Comparative Example 1 without addition, AUC last and C max It is confirmed that the levels are approximately 128% and 116.4%, respectively. That is, HyDIFFUZE TM When added, it can be seen that the diffusion of infliximab increases by about 28% compared to the case where it is not added. Moreover, in the case of Example 3, T is higher than that of Comparative Example 1. max This means that the diffusion of infliximab occurred rapidly, as the time it takes for the plasma concentration of infliximab to reach its peak after subcutaneous administration was shortened significantly.

[0081] Additionally, HyDIFFUZE reduces the content / single dose of infliximab by approximately 25%. TM In the case of Example 2, which added AUC, compared to Comparative Example 1, last and C maxIt was confirmed that the levels were approximately 104% and 94.8%, respectively. That is, HyDIFFUZE in infliximab TM When added, it was confirmed that effective efficacy was observed even when the content / single dose of infliximab was lowered.

[0082]

[0083] Example 4-15: Derivation of a subcutaneous dosage form of a pharmaceutical preparation for subcutaneous administration with excellent viscosity characteristics and stability

[0084] A subcutaneous formulation was developed to improve the stability of infliximab while having a viscosity desirable for subcutaneous administration.

[0085] Specifically, HyDIFFUZE with 120 mg / mL infliximab TM 2,000 units / mL was added, and a pharmaceutical preparation for subcutaneous administration was prepared by mixing a specific buffer, polysorbate 80 as a surfactant, sorbitol as a sugar or its derivative, a specific amino acid as a viscosity-reducing agent, and optionally a salt as a stabilizer, as in Examples 4-15 (Table 3).

[0086] In this way, it was confirmed that all of the subcutaneous pharmaceutical preparations manufactured according to Example 4-15 can store infliximab under stable conditions by maintaining a pH of 4.5-5.5.

[0087] Distinctive Infliximab Content (mg / mL) HyDIFFUZE TMContent (unit / mL) Buffer Polysorbate 80 (mg / mL) Sorbitol (mg / mL) Amino Acid (NaCl) (mg / mL) Example 41 20 2,000 10 mM Phosphate-Citrate 0.54 5 10 mM (=1.492 mg / mL) Methionine 0 Example 5 5.3 Example 6 10 mM (=1.742 mg / mL) Arginine 0 Example 7 5.3 Example 8 10 mM Succinic acid 10 mM (=1.492 mg / mL) Methionine 0 Example 9 5.3 Example 10 10 mM (=1.742 mg / mL) Arginine 0 Example 11 5.3 Example 12 10 mM MSodium Citrate 10 mM (=1.492 mg / mL) Methionine 0 Example 135.3 Example 1410 mM (=1.742 mg / mL) Arginine0 Example 155.3

[0088] (1) Measurement of viscosity of pharmaceutical preparations for subcutaneous administration

[0089] The pharmaceutical formulation for subcutaneous administration prepared according to Example 4-15 was stored under refrigerated conditions (2-8°C) for 4 weeks, and the absence of visible particles was visually observed. Thereafter, to develop a pharmaceutical formulation suitable for subcutaneous administration, the viscosity was repeatedly measured six times at 25°C using a viscometer (Rheosense, m-VROC II) (Table 4). At this time, a composition containing only 120 mg / mL infliximab was designated Comparative Example 1.

[0090] Viscosity of the preparation (mPa / s) Measurement 1 Measurement 2 Measurement 3 Measurement 4 Measurement 5 Measurement 6 Average Comparison Example 15.2275.2145.2415.2175.2735.2455.236 Example 46.8576.8026.8406.7916.8696.8326.832 Example 58.9899.0138.9388.9058.9178.9448.951 Example 610.47010.42010.33010.32010.36010.18010.347 Example 710.46010.50010.50010.46010.50010.49010.485 Example 85.6145.6215.6215.6115.6085.6015.613 Example 98.5838.2238.5548.6438.2208.4778.450 Example 105.3315.3145.3505.3465.3645.3495.342 Example 119.9609.8469.9659.9039.9629.9519.931 Example 1218.97018.85018.90019.05018.97018.96018.950Example 1313.38013.43013.44013.45013.52013.58013.467Example 1417.33017.52017.42017.81017.45017.99017.587Example 1516.31016.27016.36016.33016.38016.45016.350

[0091] As a result, in the case of Examples 4-11 using a specific buffer (phosphate-citrate or succinic acid), the viscosity could be maintained at 1.0 mPa / s to 12.0 mPa / s under 25°C temperature conditions, and thus, it can be considered desirable as a pharmaceutical preparation for subcutaneous administration. On the other hand, in the case of Examples 12-15 using sodium citrate as a buffer, the viscosity was confirmed to be excessively high compared to the case of Example 4-11, and thus a gelatin-like shape appeared. Therefore, it was confirmed that Examples 12-15 have limitations when used as a pharmaceutical preparation for subcutaneous administration, such as the possibility of tissue damage at the injection site, pain, and inappropriate distribution of the drug.

[0092] In particular, in the case of Examples 8 and 10, which omitted the mixing of salts while using a specific buffer (succinic acid), the viscosity could be maintained at 1.0 mPa / s to 6.0 mPa / s under temperature conditions of 25°C, and thus, it can be considered a desirable formulation in terms of viscosity characteristics. That is, in the case of Examples 8 and 10, HyDIFFUZE was added to infliximab. TM Despite the addition of infliximab, no gelatin-like shape was observed at all, and the viscosity was equivalent to that of Comparative Example 1 containing infliximab alone.

[0093]

[0094] (2) Measurement of infliximab content in pharmaceutical preparations for subcutaneous administration

[0095] The pharmaceutical preparation for subcutaneous administration manufactured according to Example 4-11 was stored under refrigerated conditions (2-8°C) for 0, 1, 2, 3, and 4 weeks, and then the absorbance was measured at a wavelength of around 280 nm (UV-280, Aglient Carry 100 spectrometer) to confirm the change in the content of infliximab (Table 5).

[0096] Change in infliximab content (%) Week 1 Week 2 Week 3 Week 4 Example 4 100.00104.04 89.3797.7286.94 Example 5 100.00108.9095.8098.6977.03 Example 6 100.0095.14 89.6788.8299.23 Example 7 100.0099.6087.66 108.3798.34 Example 8 100.00124.7393.14 106.0495.58 Example 9 100.00104.3490.90102.4092.08 Example 10100.00103.41111.25101.2675.02 Example 11100.00109.80108.11108.60101.20

[0097] As a result, in the case of Examples 4-11 using specific buffers (phosphate-citrate or succinic acid), no significant change was observed as the infliximab content was maintained at 80 (±5)% or more for 4 weeks, so all formulations can be considered stable preparations.

[0098] In particular, considering both viscosity characteristics and stability, Example 8, which uses a specific buffer (succinic acid), applies methionine as a viscosity-reducing agent, and omits the mixing of salts, can be considered the most suitable subcutaneous administration formulation.

[0099]

[0100] (3) Measurement of infliximab purity in pharmaceutical preparations for subcutaneous administration

[0101] The pharmaceutical preparation for subcutaneous administration manufactured according to Example 4-11 was stored under refrigerated conditions (2-8°C) for 0, 1, 2, 3, and 4 weeks, and then the purity of infliximab was analyzed using size exclusion high performance liquid chromatography (Size Exclusion HPLC) (Table 6).

[0102] Classification Infliximab Purity (%) 0 Week 1 Week 2 Week 3 Week 4 Week Example 498.7098.9599.3899.3599.37 Example 598.7798.8899.1899.2599.35 Example 698.7298.9199.2799.4799.26 Example 798.7298.8099.1399.3299.31 Example 898.8099.0099.2599.4199.22 Example 998.8698.9699.1899.2699.26 Example 1098.8499.1499.3999.4699.31 Example 1198.8499.0199.3199.2799.21

[0103] As a result, in the case of Examples 4-11 using specific buffers (phosphate-citrate or succinic acid), the purity of infliximab in all formulations was stably maintained within ±3% for 4 weeks, and thus can be considered a stable formulation.

[0104]

[0105] (4) Measurement of the potency of natural human recombinant hyaluronidase PH20 in pharmaceutical preparations for subcutaneous administration

[0106] The pharmaceutical preparation for subcutaneous administration prepared according to Example 4-11 was stored under refrigerated conditions (2-8°C) for 4 weeks, and then the change in the titer of native human recombinant hyaluronidase PH20 was analyzed using a turbidity analysis method that measures absorbance at a wavelength of 600 nm (UV-Vis spectrophotometer) (Table 7).

[0107] DistinctionHyDIFFUZE TM Change in potency (%) 0 Week 1 Week 2 Week 3 Week 4 Week Example 4 100.009 7.12 102.6 7 99.37 100.24 Example 5 100.009 4.58 107.63 103.32 97.75 Example 6 100.001 02.17 107.23 96.8 199.15 Example 7 100.009 7.73 97.80 77.31 102.40 Example 8 100.009 2.4 1101.6 8 8 5 9 8.61 Example 9 100.009 0.36 111.39 105.40 9 9.41 Example 10100.0098.23111.02101.45101.71Example 11100.0098.71106.5897.54105.85

[0108] As a result, for examples 4-11 using specific buffers (phosphate-citrate or succinic acid), all formulations did not cause relative changes in turbidity, and HyDIFFUZE TM It was found that the activity of native human recombinant hyaluronidase PH20 was not affected as the activity was maintained at 80(±5)% or more.

[0109]

[0110] To summarize what is shown in Table 4-7, infliximab plus HyDIFFUZE TMHowever, it has been confirmed that pharmaceutical formulations using phosphate-citrate or succinic acid (especially succinic acid) as a specific buffer have a viscosity desirable for subcutaneous administration, and can maintain high stability of infliximab and native human recombinant hyaluronidase PH20 enzyme activity under refrigerated conditions (2-8°C). Such pharmaceutical formulations for subcutaneous administration appear to be applicable to nonclinical and clinical applications as subcutaneous administration dosage forms.

[0111]

[0112] Experimental Example 1: Infliximab diffusion effect according to the addition of natural human recombinant hyaluronidase PH20 when the infliximab content / single dose is reduced in a subcutaneous formulation.

[0113] When the content of infliximab / single administration dose was reduced in a subcutaneous administration formulation, we sought to determine whether the infliximab diffusion effect could be maintained depending on whether or not natural human recombinant hyaluronidase PH20 was added.

[0114] First, 84 mg / kg infliximab and HyDIFFUZE TM Example 8 was prepared as a subcutaneous administration formulation containing 1,400 units / kg, and the composition of specific buffer, surfactant, sugar derivative, and viscosity reducing agent is as shown in Table 3. At this time, Example 8 was prepared by adding HyDIFFUZE to 120 mg / mL infliximab. TM Based on Example 3, a composition containing 2,000 unit / mL of infliximab, a subcutaneous formulation was developed to have a viscosity suitable for subcutaneous administration and to improve the stability of infliximab. In addition, Example 16 was prepared as a subcutaneous formulation using the same composition as Example 8, but with a 25% reduction in the infliximab content / single dose. Meanwhile, HyDIFFUZE, which uses the same composition as Example 8 but contains only 84 mg / kg of infliximab, was prepared. TM The subcutaneous administration formulation omitting the above was used as comparative example 2 (Table 8).

[0115] HyDIFFUZE Infliximab content / single dose TM Comparison of content / single dose Infliximab 120 mg / mL - Infliximab 84 mg / kg - Example 8 Infliximab 120 mg / mL HyDIFFUZE TM 2,000 unit / mL infliximab 84 mg / kg HyDIFFUZE TM 1,400 units / kgExample 16 Infliximab 90 mg / mLHyDIFFUZE TM 2,000 unit / mL infliximab 63 mg / kg HyDIFFUZE TM 1,400 units / kg

[0116] Afterwards, a single subcutaneous injection was given to SD rats, and approximately 0.5 mL of blood was collected from the jugular vein using a disposable syringe (1 mL, 25 G) at 0 hours before administration, 2 and 7 hours after administration, and 1, 2, 3, 4, 7, 10, 14, 21, and 28 days. The blood was then placed in an EDTA tube and centrifuged at 12,000 rpm for 3 minutes to separate the plasma.

[0117] To quantitatively analyze infliximab in SD rat plasma, the antibody ELISA kit (Matriks biotek, Cat. No. INF-FD-REMI) was used. Antibody standard solutions were serially diluted 1 / 3 with Assay buffer within the calibration range of 0.41–300 ng / mL, and samples were prepared by diluting them in Assay buffer so that they could be measured within the calibration range. Experiments were performed according to the ELISA kit analysis procedure, and the absorbance at 450 nm was measured using a microplate reader. Based on the analysis results, pharmacokinetic analysis was performed using the WinNonlin program (Table 9 and Fig. 2).

[0118] Distinction AUC last (㎍·hr / mL)C max (㎍ / mL)T max (hr)t 1 / 2 (hr)CL t (mL / hr / kg)V d(mL / kg)MRT(hr)Comparative Example 2Average70488.04356.457.6037.281.2263.21143.89Standard Deviation11335.3829.3813.159.700.217.1512.03N5555555Example 8Average89660.73427.4867.2045.910.9462.39149.91Standard Deviation9685.1145.3720.088.160.1111.3814.11N5555555Example 16Mean 74379.83339.0867.2040.340.8749.32147.70Standard deviation 13719.4253.3326.297.600.133.7414.48N5555555

[0119] As a result, the content / single dose of infliximab is equivalent and HyDIFFUZE TM In the case of Example 8, which is a subcutaneous administration formulation with added HyDIFFUZE TM Compared to Comparative Example 2, which is a subcutaneous administration formulation without adding AUC last and C max It is confirmed that the levels are approximately 127.2% and 120%, respectively. That is, HyDIFFUZE TM In the case of the subcutaneous formulation with added infliximab, the diffusion of infliximab was found to increase by approximately 27.2% compared to the case without it.

[0120] Additionally, HyDIFFUZE reduces the content / single dose of infliximab by approximately 25%. TM In the case of Example 16, which is a subcutaneous administration formulation with added AUC, compared to Comparative Example 2, last and C max It was confirmed that the levels were approximately 105.5% and 95%, respectively. Similarly, in the subcutaneous formulation, infliximab was added to HyDIFFUZE TM When added, it was confirmed that effective efficacy was observed even when the content / single dose of infliximab was lowered.

[0121]

[0122] Experimental Example 2: Infliximab diffusion effect depending on the addition of natural human recombinant hyaluronidase PH20 when maintaining infliximab content and increasing single dose (administration volume) in a subcutaneous administration formulation

[0123] We wanted to determine whether the diffusion effect of infliximab could be enhanced by adding natural human recombinant hyaluronidase PH20 when the single dose was increased by increasing the administration volume while maintaining the infliximab content in the subcutaneous administration formulation.

[0124] First, 84 mg / kg infliximab and HyDIFFUZE TM Example 8 was prepared as a subcutaneous administration formulation containing 1,400 unit / kg, and the composition of the specific buffer, surfactant, sugar derivative, and viscosity reducer is as shown in Table 3. In addition, infliximab and HyDIFFUZE were prepared by applying the same composition as in Example 8 but increasing the administration volume. TM Examples 17, 18, and 19 were prepared as subcutaneous formulations in which the respective single doses were increased by 2, 3, and 4 times, respectively. Meanwhile, the same composition as in Example 8 was applied, but containing only 84 mg / kg infliximab and HyDIFFUZE TM A subcutaneous administration formulation with omitted was designated as Comparative Example 2 (Table 10).

[0125] Classification Infliximab Content / Single Dose HyDIFFUZE TM Content / Single Dose Comparison Example 2 Infliximab 120 mg / mL - Infliximab 84 mg / kg - Example 8 Infliximab 120 mg / mL HyDIFFUZE TM 2,000 unit / mL infliximab 84 mg / kg HyDIFFUZE TM 1,400 unit / kg Example 17 Infliximab 120 mg / mL HyDIFFUZE TM 2,000 unit / mL infliximab 168 mg / kg HyDIFFUZE TM2,800 unit / kg Example 18 Infliximab 120 mg / mL HyDIFFUZE TM 2,000 unit / mL infliximab 252 mg / kg HyDIFFUZE TM 4,200 unit / kg Example 19 Infliximab 120 mg / mL HyDIFFUZE TM 2,000 unit / mL infliximab 336 mg / kg HyDIFFUZE TM 5,600 units / kg

[0126] Pharmacokinetic analysis was performed using the same method as in Experimental Example 1 (Table 11 and Fig. 3).

[0127] Distinction AUC last (㎍·hr / mL)C max (㎍ / mL)T max (hr)t 1 / 2 (hr)CL t (mL / hr / kg)V d (mL / kg)MRT(hr) Comparative Example 2 Average 70 488.0 435 6.4 57.6 03 7.2 81.2 26 3.2 11 43.89 Standard Deviation 11 33 35.3 29.3 81 3.1 59.7 00.2 17.1 51 2.0 3N 5555555 Example 8 Average 89 660.7 342 7.4 86 7.2 04 5.9 10.9 46 2.3 91 49.9 1 Standard Deviation 96 85.1 45.3 20.0 88.1 60.1 11 11 38 14.1 1N 5555555 Example 17 Mean 15 19 44.6 47 61.2 43 8.4 0 59.3 11.1 38 8.1 61 48.95 Standard Deviation 29 78 1.9 20 6.6 13.1 55 0.8 50.2 35 6.1 62 0.51 N5555555 Example 18 Mean 32 47 1.4 31 34 2.0 6 57.6 6 8.7 60.8 0 76.2 71 86.57 Standard Deviation 57 66 5.4 78 6.0 13.1 52 6.3 70.1 52 2.4 30.4 4 N5555555 Example 19 Average 355 10 5.95 19 10.6 25 2.8 0 38.75 1.0 5 2.5 7 14 0.28 Standard Deviation 8 8 6 8 2.6 5 5 0 7.8 7 10.7 3 12.6 4 0.2 8 9.1 7 15.36 N 5 5 5 5 5 5

[0128] As a result, as the single dose of infliximab increased, HyDIFFUZETM In the case of Examples 17, 18, and 19, which are subcutaneous formulations containing HyDIFFUZE TM Compared to Comparative Example 2, a subcutaneous administration formulation without added, AUC last and C max It is confirmed to be at levels of approximately 215.6–503.8% and approximately 214–536%. In other words, HyDIFFUZE together with high-dose infliximab TM When added, the overall uptake kinetics of infliximab increased significantly compared to when not added. Therefore, as the plasma concentration of infliximab is continuously maintained above the minimum effective concentration, the point at which it drops below the minimum effective concentration may be delayed, and consequently, this suggests that the administration interval of the subcutaneous formulation can be increased or the number of administrations reduced (Table 11 and Fig. 3).

[0129]

[0130] Experimental Example 3: Substitution effect of intravenous administration by adding natural human recombinant hyaluronidase PH20 when maintaining infliximab content and increasing single-administration dose (administration volume) in a subcutaneous administration formulation

[0131] We aimed to determine whether the addition of native human recombinant hyaluronidase PH20 could replace intravenous administration when the single-dose dose was increased by increasing the administration volume while maintaining the infliximab content in the subcutaneous administration formulation.

[0132] First, 168 mg / kg infliximab and HyDIFFUZE TM Example 17 was prepared as a subcutaneous administration formulation containing 2,800 units / kg, and the composition of specific buffers, surfactants, sugar derivatives, and viscosity-reducing agents was referred to Table 3. The same subcutaneous administration formulation composition as Example 17 was applied, but the administration volume was increased to include infliximab and HyDIFFUZE. TMExample 18 was prepared as a subcutaneous administration formulation in which the respective single dose was increased by 1.5 times. Meanwhile, the same composition as in Examples 17 and 18 was applied, but containing only 60 mg / kg infliximab and HyDIFFUZE TM An intravenous (IV) formulation with the omission of [the part] was designated as Comparative Example 3 (IV) (Table 12).

[0133] Classification Infliximab Content / Single Dose HyDIFFUZE TM Content / Single Dose Comparison Example 3 (IV) Infliximab 10 mg / mL - Infliximab 60 mg / kg - Example 17 Infliximab 120 mg / mL HyDIFFUZE TM 2,000 unit / mL infliximab 168 mg / kg HyDIFFUZE TM 2,800 unit / kg Example 18 Infliximab 120 mg / mL HyDIFFUZE TM 2,000 unit / mL infliximab 252 mg / kg HyDIFFUZE TM 4,200 unit / kg

[0134] Pharmacokinetic analysis was performed using the same method as in Experimental Example 1 (Table 13 and Fig. 4).

[0135] Distinction AUC last (㎍·hr / mL)C max (㎍ / mL)T max (hr)t 1 / 2 (hr)CL t (mL / hr / kg)V d(mL / kg)MRT(hr)Comparative Example 3(IV)Average168764.101373.66444.950.3422.47223.38Standard Deviation9263.77148.662.737.720.024.0211.25N5555555Example 17Average151944.64761.2438.4059.311.1388.16148.95Standard Deviation29781.90206.6613.1550.850.2356.1620.51N5555555Example 18 Average 320 47 1.43 134 2.06 57.66 8.76 0.80 76.27 186.57 Standard Deviation 57 66 5.47 86.07 13.15 26.37 0.15 22.41 30.44 N 5555555

[0136] As a result, as the single dose of infliximab increased, HyDIFFUZE TM In the case of Examples 17 and 18 with added HyDIFFUZE, despite being a subcutaneous administration formulation, TM Compared to Comparative Example 3, an intravenous formulation without added, AUC last and C max It is confirmed to be at levels of approximately 90-189.9% and 55.4-97.7%. In other words, HyDIFFUZE together with high-dose infliximab TM When administered subcutaneously with the addition of [the substance], it can be considered to have an effective therapeutic effect equivalent to that of intravenous administration.

[0137]

[0138] The foregoing description of the present invention is for illustrative purposes only, and those skilled in the art will understand that other specific forms can be easily modified without altering the technical spirit or essential features of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

Claims

1. Infliximab; and A pharmaceutical preparation for subcutaneous administration comprising a natural human recombinant hyaluronidase PH20.

2. In paragraph 1, A pharmaceutical preparation for subcutaneous administration, characterized in that the above-mentioned natural human recombinant hyaluronidase PH20 is used for diffusion of the above-mentioned infliximab.

3. In paragraph 1, A pharmaceutical preparation for subcutaneous administration, characterized in that the content of the above infliximab is 30 mg / mL to 240 mg / mL, and the content of the above natural human recombinant hyaluronidase PH20 is 1,000 unit / mL to 10,000 unit / mL.

4. In paragraph 1, A pharmaceutical preparation for subcutaneous administration, characterized in that when the above pharmaceutical preparation for subcutaneous administration is administered subcutaneously, the single administration dose of infliximab is 30 mg to 600 mg.

5. In paragraph 1, A pharmaceutical preparation for subcutaneous administration, characterized in that when the above pharmaceutical preparation for subcutaneous administration is administered subcutaneously, the area under the concentration-time curve (AUC) of infliximab in plasma is 120% or more compared to when a preparation containing only infliximab is subcutaneously administered at the same dose.

6. In paragraph 1, A pharmaceutical preparation for subcutaneous administration, characterized in that it additionally contains a phosphate buffer or a succinic acid buffer as a buffer.

7. In paragraph 1, A pharmaceutical preparation for subcutaneous administration, characterized in that it additionally contains 1 mM to 20 mM succinic acid as a buffer.

8. In paragraph 1, A pharmaceutical preparation for subcutaneous administration, characterized in that the above pharmaceutical preparation for subcutaneous administration excludes salt.

9. In paragraph 1, As a surfactant, a polysorbate-based substance; As a sugar or a derivative thereof, at least one selected from the group consisting of sorbitol, mannitol, trehalose and sucrose; and A pharmaceutical preparation for subcutaneous administration, characterized in that it additionally contains methionine or arginine as a viscosity-reducing agent.

10. In paragraph 1, As a surfactant, polysorbate 80 at 0.05 mg / mL to 5 mg / mL; As a sugar or its derivative, sorbitol of 10 mg / mL to 100 mg / mL; and A pharmaceutical preparation for subcutaneous administration, characterized in that it additionally contains 0.5 mg / mL to 20 mg / mL of methionine or arginine as a viscosity-reducing agent.

11. In paragraph 1, The above pharmaceutical preparation for subcutaneous administration is characterized in that the viscosity is 1.0 mPa / s to 12.0 mPa / s under temperature conditions of 25°C.

Images